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Updated: Mar 8, 2026
"Cell Surface Capture" Workflow for Label-Free Quantification of the Cell Surface Proteome
Published on: March 24, 2023
Improved surfaceome coverage with a label-free nonaffinity-purified workflow
Tina Glisovic-Aplenc1,2, Saar Gill3, Lynn A Spruce4
1Division of Oncology, Center for Childhood Cancer Research, The Children's Hospital of Philadelphia, Philadelphia, PA, USA.
Abstract:
The proteins of the cellular plasma membrane (PM) perform important functions relating to homeostasis and intercellular communication. Due to its overall low cellular abundance, amphipathic character, and low membrane-to-cytoplasm ratio, the PM proteome has been challenging to isolate and characterize, and is poorly represented in standard LC-MS/MS analyses. In this study, we employ sucrose gradient ultracentrifugation for the enrichment of the PM proteome, without chemical labeling and affinity purification, together with GeLCMS and use subsequent bioinformatics tools to select proteins associated with the PM/cell surface, herein referred to as the surfaceome. Using this methodology, we identify over 1900 cell surface associated proteins in a human acute myeloid leukemia cell line. These surface proteins comprise almost 50% of all detected cellular proteins, a number that substantially exceeds the depth of coverage in previously published studies describing the leukemia surfaceome.
Insights
Researchers identified over 1900 cell surface proteins in leukemia cells, significantly expanding the known surfaceome. This breakthrough improves understanding of plasma membrane proteins and their roles in cellular functions.
Area of Science:
- Proteomics
- Cell Biology
- Biochemistry
Background:
- Plasma membrane (PM) proteins are crucial for cell homeostasis and communication.
- The PM proteome is difficult to study due to low abundance and challenging isolation.
- Standard methods often underrepresent PM proteins in analyses.
Purpose of the Study:
- To develop an effective method for enriching and characterizing the plasma membrane proteome.
- To identify a comprehensive set of cell surface proteins (surfaceome) in leukemia cells.
- To overcome limitations of previous studies on the leukemia surfaceome.
Main Methods:
- Utilized sucrose gradient ultracentrifugation for plasma membrane proteome enrichment.
- Employed Gel-liquid chromatography-tandem mass spectrometry (GeLC-MS) for protein identification.
- Applied bioinformatics tools to specifically select for PM/cell surface-associated proteins.
Main Results:
- Identified over 1900 cell surface-associated proteins in a human acute myeloid leukemia cell line.
- The identified surfaceome constitutes nearly 50% of all detected cellular proteins.
- This represents a substantial increase in coverage compared to previous leukemia surfaceome studies.
Conclusions:
- The developed method effectively enriches and characterizes the plasma membrane proteome.
- This study significantly expands the catalog of known leukemia surface proteins.
- The findings provide a deeper understanding of the leukemia surfaceome and its potential roles.
